Altered Lymphatic Function and Development in Congenital Heart Disease

先天性心脏病中淋巴功能和发育的改变

• 批准号: 10590656
• 负责人: Sanjeev A. Datar
• 金额: $ 72.86万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-06-01 至 2027-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10590656
• 关键词: Adult; Animal Model; Animals; Antioxidants; Architecture; Biological Availability; Blood flow; Cannulations; Cardiac; Catheters; Cell Culture Techniques; Cell Proliferation; Cell Separation; Characteristics; Child; Chronic; Chylothorax; Devices; Disease; Endothelin-1; Exposure to; Genetic Transcription; Heart Abnormalities; Hypoxia Inducible Factor; Immunologics; In Vitro; Lung; Lung diseases; Lymphatic; Lymphatic Endothelial Cells; Lymphatic Endothelium; Lymphatic clearance; Lymphatic function; Magnetic Resonance Imaging; Mediating; Membrane Potentials; Metabolic; Mitochondria; Molecular; Morbidity - disease rate; Nitric Oxide; Pathogenesis; Patients; Phenotype; Physiological; Piezo 1 ion channel; Play; Pneumonectomy; Prevention strategy; Proliferating; Pulmonary function tests; Reactive Oxygen Species; Respiration Disorders; Respiratory Mechanics; Risk Factors; Role; SRC gene; Shunt Device; Signal Pathway; Signal Transduction; Source; Structure; Testing; Thoracic Duct; Time; Translating; antagonist; bosentan; cell growth; clinically relevant; congenital heart disorder; data integration; endothelial dysfunction; experimental study; genetic manipulation; hemodynamics; improved; in vivo; in vivo evaluation; lymph flow; lymphatic development; lymphatic dysfunction; lymphatic malformations; mechanical force; metabolic abnormality assessment; mitochondrial membrane; mitoquinone; mortality; novel; pharmacologic; programs; receptor; repaired; shear stress; therapeutic target; treatment strategy; vascular abnormality

PROJECT SUMMARY / ABSTRACT Aberrations in lymphatic structure and function are increasingly recognized as a significant source of morbidity in a variety of disease states. For example, lymphatic abnormalities associated with congenital heart disease (CHD) that result in increased pulmonary blood flow (PBF) include congenital or acquired chylothoraces, and immunologic aberrations. Importantly, it is increasingly appreciated that lymphatic abnormalities are not only associated with a variety of pulmonary diseases but participate in their pathogenesis. Recent recognition that abnormal respiratory dysfunction often persists years after cardiac correction and is an independent risk factor for mortality in adult CHD patients has brought renewed urgency to better understand the underlying lymphatic pathobiology in CHD, which thus far remains largely unknown. We have previously used a clinically relevant large animal model of CHD with increased PBF (shunt) in combination with isolated vessel reactivity of the thoracic duct and primary lymphatic endothelial cell (LEC) culture, to demonstrate that chronically increased PBF and the consequential increase in lymphatic flow, is associated with: 1) abnormal pulmonary lymphatic flow and architecture; 2) increased mitochondrial reactive oxygen species (mtROS)-driven hypoxia inducible factor-1 (HIF-1) activity and metabolic reprograming to support cellular and proliferation; and 3) a KLF2-dependent decrease in nitric oxide (NO) signaling. In this renewal application we will test our novel hypothesis that in the setting of increased PBF, the mechanosensory channel Piezo1 plays a pivotal role sensing alterations in lymphatic flow, triggering downstream increases in endothelin-1 (ET-1) and mtROS driven HIF-1a activity. This results in decreased NO bioavailability and subsequent lymphatic dysfunction, that contributes to persistently abnormal respiratory mechanics even after the cardiac defect has been repaired. In support of this hypothesis, we present preliminary evidence demonstrating: 1) increased lymphatic endothelial Piezo1, ET-1, RhoA, and mtROS in shunt LECs; 2) Piezo1-, ET-1-, and mtROS-dependent increases in HIF-1 in control LECs; 3) shunt lambs have abnormal respiratory mechanics and lymphatic endothelial dysfunction that persists following closure of the shunt, despite normalized hemodynamics, and 4) that treatment with a mitochondrially-targeted antioxidant (mitoquinone, MitoQ) reverses the HIF-1 mediated shunt LEC phenotype in vitro and normalizes pulmonary lymphatic architecture and function in vivo. This overall hypothesis will be tested in three inter-related, but independent mechanistic aims, that utilize integrated physiologic, cellular, and molecular experiments. These translational and targetable studies include: 1) whole animal hemodynamic physiologic studies, advanced CT/MR imaging, and sophisticated pulmonary function testing, 2) ex vivo thoracic duct reactivity studies, and 3) transcriptional and metabolic studies in LECs. A deeper understanding of the mechanisms investigated may lead to improved treatment and prevention strategies for lymphatic abnormalities in the setting of CHD and other disease states, including pneumonectomy and other vascular abnormalities that result in increased PBF.

项目摘要 /摘要 淋巴结构和功能的畸变越来越多地被认为是发病率的重要来源 在各种疾病状态下。例如，与先天性心脏病相关的淋巴异常 （CHD）导致肺血流增加（PBF）包括先天性或熟悉的乳房，以及 免疫畸变。重要的是，越来越多地赞赏淋巴异常不仅是 与多种肺部疾病有关，但参与了它们的发病机理。最近的认可 心脏矫正后数年几年，呼吸障碍异常，是一个独立的危险因素 对于成年冠心病患者的死亡率，已经带来了新的紧迫性，以更好地了解潜在的淋巴管 冠心病中的病理生物学，到目前为止，这仍然是未知的。我们以前已经使用了临床上相关的 冠心病的大型动物模型与PBF（分流）增加，并结合孤立的血管反应性 胸腔导管和原发性淋巴内皮细胞（LEC）培养，以证明长期增加了PBF 淋巴流的结果增加与：1）异常的肺淋巴流动和 建筑学; 2）增加线粒体活性氧（MTROS）驱动的缺氧诱导因子1 （HIF-1）活性和代谢重编程，以支持细胞和增殖； 3）KLF2依赖性 一氧化氮（NO）信号传导的减少。在此续订应用中，我们将测试我们的新假设 设置PBF的设置，机械学通道Piezo1扮演着关键的角色传感改变 淋巴流，触发下游的淋巴流量增加了内皮素-1（ET-1）和MTROS驱动的HIF-1A活性。这 导致没有任何生物利用度和随后的淋巴功能障碍，这有助于持续 即使修复心脏缺陷后，呼吸力学异常。为了支持这一假设， 我们提供了证明：1）淋巴内皮压电1，ET-1，RHOA和RHOA和RHOA和 分流LEC中的Mtros； 2）对照LEC中HIF-1的Piezo1-，ET-1-和MTROS依赖性增加； 3）分流 羔羊具有闭合后持续存在的异常呼吸力学和淋巴内皮功能障碍 分流的，dospite标准化血液动力学，4）用线粒体靶向抗氧化剂进行治疗 （Mitoquinone，mitoq）在体外逆转HIF-1介导的分流LEC表型，并归一化肺 体内淋巴结构和功能。该总体假设将在三个相互关联的情况下进行测试，但 独立的机械目的，利用综合的生理，细胞和分子实验。这些 翻译和目标研究包括：1）全动物血液动力学生理研究，先进 CT/MR成像和复杂的肺功能测试，2）离体胸腔导管反应性研究，3） LEC中的转录和代谢研究。对研究机制的更深入了解可能会导致 改善冠心病和其他环境中淋巴异常的治疗和预防策略 疾病状态，包括肺部切除术和其他血管异常，导致PBF增加。

项目成果

Respiratory mechanics and gas exchange in an ovine model of congenital heart disease with increased pulmonary blood flow and pressure.

• DOI: 10.3389/fphys.2023.1188824
• 发表时间: 2023
• 期刊: Frontiers in physiology
• 影响因子: 4
• 作者: Soares JHN;Raff GW;Fineman JR;Datar SA
• 通讯作者: Datar SA

Preservation of myocardial contractility during acute hypoxia with OMX-CV, a novel oxygen delivery biotherapeutic.

• DOI: 10.1371/journal.pbio.2005924
• 发表时间: 2018-10
• 期刊: PLoS biology
• 影响因子: 9.8
• 作者: Boehme J;Le Moan N;Kameny RJ;Loucks A;Johengen MJ;Lesneski AL;Gong W;Goudy BD;Davis T;Tanaka K;Davis A;He Y;Long-Boyle J;Ivaturi V;Gobburu JVS;Winger JA;Cary SP;Datar SA;Fineman JR;Krtolica A;Maltepe E
• 通讯作者: Maltepe E